Figure 9.

DNA Binding Is Not Necessary for the Light-Induced Degradation of PIF1.

(A) The PIF1E293D mutant does not bind to a G-box DNA sequence element (POR C; Su et al., 2001; Moon et al., 2008). In vitro translated PIF1 or PIF1E293D was incubated with a radiolabeled fragment of POR C in a DNA gel shift assay. Lane 1, free probe; lanes 2 and 3, increasing amounts of wild-type PIF1; lanes 4 and 5, increasing amounts of PIF1E293D mutant protein; lane 6, unrelated luciferase protein as a negative control. FP, free probes.

(B) Comparison of the levels of wild-type and mutant PIF1 proteins produced by in vitro transcription and translation.

(C) Relative LUC assays were performed under the conditions described for Figure 8. Means ± se of five biological replicates are shown.